Day 3: Difference between revisions
New page: 2. Outcome #1, no colonies on any plate: This would suggest that the transformation was unsuccessful, because even the positive control with the un-engineered M13K07 DNA failed to grow in... |
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2. | 2.Outcome #1, no colonies on any plate: This would suggest that the transformation was unsuccessful, because even the positive control with the un-engineered M13K07 DNA failed to grow in the presence of kanamycin, meaning the DNA was not taken up by the cell. | ||
Outcome #1, no colonies on any plate: This would suggest that the transformation was unsuccessful, because even the positive control with the un-engineered M13K07 DNA failed to grow in the presence of kanamycin, meaning the DNA was not taken up by the cell. | |||
Outcome #2, thousands of colonies on all plates: This could indicate contamination because the cells transformed with the negative control for ligation, backbone only and no ligase, should not have kanamycin resistance. | Outcome #2, thousands of colonies on all plates: This could indicate contamination because the cells transformed with the negative control for ligation, backbone only and no ligase, should not have kanamycin resistance. | ||
Outcome #3, approximately the same number of colonies on the backbone+ligase+kill cut as the backbone+insert+ligase+kill cut: This could also indicate that there wasn't complete digestion during the kill-cut and some backbone managed to back-ligate and be transformed into the cells of both samples. | Outcome #3, approximately the same number of colonies on the backbone+ligase+kill cut as the backbone+insert+ligase+kill cut: This could also indicate that there wasn't complete digestion during the kill-cut and some backbone managed to back-ligate and be transformed into the cells of both samples. | ||
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! Diagnostic digest 2 | |||
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Revision as of 20:34, 20 September 2007
2.Outcome #1, no colonies on any plate: This would suggest that the transformation was unsuccessful, because even the positive control with the un-engineered M13K07 DNA failed to grow in the presence of kanamycin, meaning the DNA was not taken up by the cell.
Outcome #2, thousands of colonies on all plates: This could indicate contamination because the cells transformed with the negative control for ligation, backbone only and no ligase, should not have kanamycin resistance.
Outcome #3, approximately the same number of colonies on the backbone+ligase+kill cut as the backbone+insert+ligase+kill cut: This could also indicate that there wasn't complete digestion during the kill-cut and some backbone managed to back-ligate and be transformed into the cells of both samples.
3.
| Diagnostic digest 1 | plasmid with insert | plasmid no insert |
|---|---|---|
| Enzyme(s) used | ||
| Buffer used | ||
| Temperature | ||
| Predicted fragments | ||
| Diagnostic digest 2 | ||
| Enzyme(s) used | ||
| Buffer used | ||
| Temperature | ||
| Predicted fragments |